Metallization of Bacterial Cellulose for Electrical and Electronic Device Manufacture

ABSTRACT

A method for the deposition of metals in bacterial cellulose and for the employment of the metallized bacterial cellulose in the construction of fuel cells and other electronic devices is disclosed. The method for impregnating bacterial cellulose with a metal comprises placing a bacterial cellulose matrix in a solution of a metal salt such that the metal salt is reduced to metallic form and the metal precipitates in or on the matrix. The method for the construction of a fuel cell comprises placing a hydrated bacterial cellulose support structure in a solution of a metal salt such that the metal precipitates in or on the support structure, inserting contact wires into two pieces of the metal impregnated support structure, placing the two pieces of metal impregnated support structure on opposite sides of a layer of hydrated bacterial cellulose, and dehydrating the three layer structure to create a fuel cell.

CROSS-REFERENCES TO RELATED APPLICATIONS

This is a divisional application of application Ser. No. 11/207,431filed Aug. 19, 2005, which is a divisional application of applicationSer. No. 10/017,202 filed Dec. 14, 2001, now U.S. Pat. No. 6,986,963.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

This invention was made with Government support under Contract No.DE-AC05-000R22725 awarded to UT-Battelle, LLC, by the U.S. Department ofEnergy. The Government has certain rights in this invention.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to a method for the deposition of metals inbacterial cellulose and the utilization of the metallized bacterialcellulose in the construction of fuel cells and other electronicdevices.

2. Description of the Related Art

The production of bacterial cellulose (also referred to as microbialcellulose) from cellulose-synthesizing bacterium has been studied forover half a century. For instance, it was reported back in 1947 that inthe presence of glucose and oxygen, resting cells of Acetobacter xylinumsynthesize cellulose (see, S. Hestrin et al., “Synthesis of Cellulose byResting Cells of Acetobacter xylinum”, Nature 159: 64-65, 1947).

Through subsequent studies, it was determined that the physicalproperties of bacterial cellulose differ from those of the celluloseproduced by green plants. Upon visual examination, it is evident thatplant and bacterial cellulose differ in appearance and water content.Plant cellulose has a fibrous structure, while bacterial celluloseresembles a gel. In its hydrated state, the bacterial cellulose containsover a hundred times its weight in water. Yet both of these substancesare built from the same basic unit, chains of glucose molecules that arelinked by β-1,4-glycosidic bonds. The difference in the properties ofthese materials results from their nanoscale structural architecture.Cellulose that is synthesized by plants such as cotton (Gossypium spp.)and ramie (Boehmeria nivea) has a structure resembling a heavy-duty ropemade of many small fibers twisted into larger fibers that are thentwisted into the rope. Thirty-six glucose chains are assembled into anelementary fibril with a diameter of 3.5 nanometers. Microfibrils areassembled into macrofibrils that have a diameter ranging from 30 to 360nanometers. The macrofibrils are then assembled into fibers. Imaging ofcotton linter fibers by atomic force microscopy found an averagemacrofibril diameter of approximately 100 nanometers (see, Hon,“Cellulose: a random walk along its historical path”, Cellulose 1:1-251994; and Franz et al. “Cellulose”, in Methods in Plant Biochem. Vol. 2,Chapter 8, P. M. Dey and J. B. Harborne, editors, Academic Press,London, pages 291-322, 1990).

The most widely studied cellulose-synthesizing bacterium is Acetobacterxylinus (formerly Acetobacter xylinum, recently renamedGluconacetobacter xylinus according to the American Type CultureCollection). In fact, this microorganism has been used for theproduction of the food product nata de coco in the Philippines.Cellulose is secreted by Acetobacter in the form of a twisted ribbon 40to 60 nanometers wide that is extruded at a rate of 2micrometers/minute. Each ribbon consists of 46 microfibrils, each ofwhich has an average cross-section of 1.6×5.8 nanometers. These twistedribbons, roughly corresponding to the macrofibrils of plant cellulose,assemble into sheets outside the cell, that combine to form acentimeter-thick layer called a pellicule on the surface of the culturemedium. Scanning electron microscopy has revealed that, inside thepellicule, the fibrils are organized to form tunnels with a diameter of7 micrometers, large enough for the bacteria to move through (see, S.Hestrin et al., “Synthesis of Cellulose by Resting Cells of Acetobacterxylinum”, Nature 159: 64-65, 1947; S. Hestrin, et al., “Synthesis ofcellulose by Acetobacter xylinum: Preparation of freeze-dried cellscapable of polymerizing glucose to cellulose”, Biochem. J. 58: 345-352,1954; and Cannon et al., “Biogenesis of Bacterial Cellulose”, Crit.Reviews in Microbiol. 17(6): 435-447, 1991).

The aforementioned nata de coco or coconut gel has been produced fordomestic consumption in the Philippines for at least 100 years. Nata decoco is the gel-like cellulose pellicule formed on the surface of mediaby Acetobacter xylinum cultures. In recent years, it has become one ofthe most popular Filipino food exports. The export of nata de coco grewfrom $1.0 million in 1992 to $25.9 million in 1993, with 95% of thetotal going to Japan. Traditional production of nata de coco is carriedout in the Philippines as a cottage industry. Fermentation of coconutmilk and glucose medium inoculated with starter is carried out understatic culture conditions, i.e., in square plastic containers 1.5centimeters high. The fermentation broth is acidified by the addition ofacetic acid. Typically, a fermentation time of 10 to 12 days at ambienttemperature is required for production of a layer or pellicule 1centimeter thick. The pellicules are washed with water and, in somecases, sodium hydroxide solution, then cut into 1 centimeter cubes. Thecubes are generally soaked in sucrose solutions with addition offlavorings and colors for the food product (see, Technology andLivelihood Resource Center, Makati City, Philippines, available on theInternet at http://esprint.com.ph/cocosoy/mainpage/mainpage.html).

The unique properties of the bacterial cellulose synthesized byAcetobacter have inspired attempts to use it in a number of commercialproducts. These include tires (see, e.g., U.S. Pat. No. 5,290,830),headphone membranes (see, e.g., U.S. Pat. No. 4,742,164), paper (see,e.g., U.S. Pat. No. 4,863,565), and textiles (see, e.g., U.S. Pat. No.4,919,753). Medical applications include a specially prepared membraneto be used as a temporary skin substitute for patients with large burnsor ulcers (see, e.g., U.S. Pat. No. 4,912,049, and Fontana, et al.,“Acetobacter Cellulose Pellicule as a Temporary Skin Substitute”, Appl.Biochem. Biotech. 24/25: 253-264 12, 1990). A patent has also issued onthe possible use of bacterial cellulose preparations as a source ofdietary fiber (see, e.g., U.S. Pat. No. 4,960,763). Numerous patentshave issued on the production of bacterial cellulose modified in somemanner during cell growth or during processing (see, e.g., U.S. Pat.Nos. 5,079,162, 5,871,978, 6,060,289, 5,955,326, 5,962,277, 5,962,278,6,017,740, and 6,071,727). It has also been reported that the additionof certain dyes to the culture medium inhibits the assembly of thepellicule sheets (see, Brown et al., “Experimental Induction of AlteredNonmicrofibrillar Cellulose”, Science 218: 1141-1142, 1982), and thatthe addition of carboxymethylcellulose to the medium results in theformation of cellulose with special optical properties (see, e.g., U.S.Pat. No. 4,942,128).

In addition to studies directed to bacterial cellulose and it uses,others have studied the chemical reactions of cellulose more generally.For instance, various mechanisms of cellulose hydrolysis has beenstudied and reported on by Lassig, Shultz, Gooch, Evans and Woodward in“Inhibition of Cellobiohydrolase I from Trichoderma reesei byPalladium”, Arch. Biochem. Biophys. 322: 119-126, 1995, and in“Palladium—a new inhibitor of cellulase activity”, Biochem. Biophys.Res. Comm. 209: 1046-1052, 1995. As part of research studying themechanism of cellulose hydrolysis, several metal ions and complexes weretested for inhibition of cellobiohydrolase I (CBH I), the β-1,4-glucanhydrolase comprising the major component of the cellulase mixturesecreted by the fungus Trichoderma reesei. The most importantcontribution of CBH I to the hydrolysis of crystalline celluloseappeared to be the binding and disruption of the cellulose fibers.Specifically, a compound was sought that would inhibit hydrolysis of theβ-1,4-glycosidic bond of the cellulose chains but would not effectbinding to the crystalline cellulose. Sodium and ammoniumhexachloropalladate were found to be the most effective inhibitors ofthe compounds screened. The hexachloropalladate inhibited hydrolysis ofboth small soluble substrates and crystalline, insoluble cellulose byCBH I, but did not inhibit binding of the enzyme to the insolublecellulose.

For years, cellulose-containing products in general have also beenstudied for uses as an alternative source of fuel. For instance, theconversion of biomass to energy has been studied for some time.Increased use of cellulose-containing products in heat or electricity(power) generation systems would be particularly desirable as productswhich contain cellulose are a renewable resource. However, the use ofbacterial cellulose in power generation systems has not beeninvestigated.

One power generation system that has attracted widespread interest isthe fuel cell. There are different types of fuel cells, but they eachproduce electrical energy by means of chemical reaction. One type offuel cell is the polymer electrolyte membrane fuel cell which comprisesa polymeric electrolyte membrane sandwiched between an anode and acathode. The fuel cell generates electrical power by bringing a fuelinto contact with the anode and an oxidant into contact with thecathode. The fuel is typically a hydrogen-containing material (forexample, water, methane, methanol or pure hydrogen), and may be suppliedto the fuel cell in liquid form or gaseous form, such as hydrogen gas.The fuel is introduced at the anode where the fuel reactselectrochemically in the presence of a catalyst to produce electrons andprotons in the anode. The electrons are circulated from the anode to thecathode through an electrical circuit connecting the anode and thecathode. Protons pass through the electrolyte membrane (which is anelectron insulator and keeps the fuel and the oxidant separate) to thecathode. Simultaneously, an oxygen-containing oxidant, such as oxygengas or air, is introduced to the cathode where the oxidant reactselectrochemically in the presence of a catalyst consuming the electronscirculated through the electrical circuit and the protons at thecathode. The halfcell reactions at the anode and the cathode are,respectively: H₂→>2H⁺+2e⁻ and ½O₂+2H⁺+2e⁻→H₂O. The external electricalcircuit withdraws electrical current and thus receives electrical powerfrom the cell. The overall fuel cell reaction produces electrical energywhich is the sum of the separate halfcell reactions written above.

While fuel cells are highly efficient electrochemical energy conversiondevices that directly convert the chemical energy derived from renewablefuels into electrical energy, they do have disadvantages. Specifically,long felt needs generally exist to reduce initial costs and provide forinexpensive maintenance of fuel cell installations. The high cost offabricating the fuel cells is due to many factors including (among otherthings) the high cost of synthetic polymeric materials used in theelectrodes and the electrolyte membranes, the safety and environmentalmeasures necessary for safe manufacture of the electrodes andelectrolyte membranes, the difficulties in controlling the concentrationof expensive catalysts, and the problems associated with the bonding ofelectrodes and the electrolyte membrane. Furthermore, it can be quiteexpensive to replace worn out fuel cells and recover the anode andcathode catalysts that may become fouled during operation. Also, fuelcell materials may not be amenable to recycling because of the presenceof metal catalysts.

Therefore, there is a need for a fuel cell power generation system thatcan be fabricated from components that are inexpensive to manufactureand that can be readily recycled and recovered. In particular, it wouldbe beneficial if these fuel cell components could be manufactured from arenewable resource such as a natural cellulose containing material.

SUMMARY OF THE INVENTION

The foregoing needs are met by a method according to the invention forthe deposition of metals in bacterial cellulose and for the employmentof the metallized bacterial cellulose in the construction of fuel cellsand other electronic devices. In one aspect of the invention, there isprovided a method for impregnating bacterial cellulose with a metalcomprising placing a bacterial cellulose matrix in a solution of a metalsalt for a sufficient time period such that the metal salt is reduced tometallic form and the metal precipitates in or on the matrix. The metalsalt may be a coordination compound including a transition metal complexion, and preferably, the metal salt is a coordination compound includinga platinum metal group complex ion.

In a second aspect on the invention, there is provided a method for theconstruction of a fuel cell. In a first part of the method, a hydratedbacterial cellulose support structure is placed in a solution of a metalsalt for a sufficient time period such that the metal salt is reduced tometallic form and the metal precipitates in or on the support structure.Typically, the metal salt is a platinum group metal coordinationcompound, such as a palladium coordination compound, and theconcentration of metal in the support structure is controlled by theresidence time of the support structure in the metal salt solution.Suitable contact wires are inserted in two pieces of the metalimpregnated support structure, and the two pieces of metal impregnatedsupport structure are then placed on opposite sides of a layer ofhydrated bacterial cellulose. The three layer structure is thendehydrated to create a fuel cell. The resulting fuel cell has adehydrated metal impregnated bacterial cellulose anode, a dehydratedbacterial cellulose electrolyte layer, and a dehydrated metalimpregnated bacterial cellulose cathode. When the contact wires of thefuel cell are connected to an electrical circuit and the anode iscontacted with a hydrogen-containing fuel, the fuel cell generateselectricity as measured in the circuit. It has been discovered that theelectrical current generated can be increased by pretreating the layerof bacterial cellulose forming the electrolyte layer with a metal saltand/or a sulfonated polymer before dehydration.

In a third aspect of the invention, there is provided a fuel cellelectrode comprising a support structure comprising bacterial celluloseand a transition metal catalyst disposed in or on the support structure.In a fourth aspect of the invention, there is provided a method forrecovering the catalyst from the fuel cell electrode comprising burningor hydrolyzing the support structure. In a fifth aspect of theinvention, there is provided an electrolyte membrane for a fuel cellwherein the electrolyte membrane comprises a support structurecomprising bacterial cellulose and a metal salt disposed in or on thesupport structure. In a sixth aspect of the invention, there is providedan electrolyte membrane for a fuel cell wherein the electrolyte membranecomprises a support structure comprising bacterial cellulose and asulfonated polymer disposed in or on the support structure. In a seventhaspect of the invention, there is provided an enzyme electrodecomprising a support structure comprising bacterial cellulose, acatalyst disposed in or on the support structure, and an enzyme disposedin or on the support structure.

It is therefore an advantage of the present invention to provide amethod for the deposition of metals in bacterial cellulose and for theemployment of the metallized bacterial cellulose in the construction offuel cells and other electronic devices.

It is another advantage of the present invention to provide fuel cellelectrodes and fuel cell solid electrolyte membranes that are formedfrom low cost precursors.

It is a further advantage of the present invention to provide fuel cellelectrodes and fuel cell solid electrolyte membranes that are formedusing low toxicity precursors, and low temperature and environmentallyfriendly processes.

It is yet another advantage of the present invention to provide a methodfor forming fuel cell electrodes and fuel cell solid electrolytemembranes that allows for greater control of the concentration ofcatalysts in the fuel cell electrodes and fuel cell solid electrolytemembranes.

It is still another advantage of the present invention to provide fuelcell electrodes and fuel cell solid electrolyte membranes including atleast one catalyst wherein the catalysts may be easily recovered fromthe fuel cell electrodes and fuel cell solid electrolyte membranes.

It is a still further advantage of the present invention to providelightweight fuel cell electrodes and lightweight fuel cell solidelectrolyte membranes that may be assembled together without adhesivesor glues.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features, aspects, and advantages of the presentinvention will become better understood upon consideration of thefollowing detailed description, appended claims, and drawings where:

FIG. 1A is a schematic end view of a metallized cellulose cube havingcontact wires inserted therein;

FIG. 1B is a schematic perspective view of the metallized cellulose cubeof FIG. 1A having the contact wires inserted therein;

FIG. 2 is a schematic side view of a fuel cell assembled from layers ofmetallized bacterial cellulose;

FIG. 3 is a schematic drawing of a fuel cell assembly used for testingthe fuel cell of FIG. 2;

FIG. 4 is a graph showing the current versus time performance of a fuelcell having metallized bacterial cellulose electrodes separated by anuntreated bacterial cellulose electrolyte layer, the fuel cell usingacid displacement as the hydrogen source;

FIG. 5 is a graph showing the current versus time performance of a fuelcell having metallized bacterial cellulose electrodes separated bypotassium chloride treated bacterial cellulose electrolyte layer, thefuel cell using acid displacement as the hydrogen source; and

FIG. 6 is a graph showing the voltage versus current performance of afuel cell having metallized bacterial cellulose electrodes separated bypotassium chloride treated bacterial cellulose electrolyte layer, thefuel cell using acid displacement as the hydrogen source.

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect, the present invention provides a method for thedeposition of metals in bacterial cellulose. The bacterial celluloseused in the method can be a bacterial cellulose matrix obtained by knownmethods. For example, the bacterial cellulose can be the Philippine foodproduct nata de coco that is produced from coconut milk and sucrose byfermentation with a starter culture containing the bacteriumGluconoacetobacter xylinus (formerly Acetobacter xylinum) or relatedspecies. Alternatively, the bacterial cellulose can be produced bygrowing Gluconoacetobacter xylinus on media formulations that have beenreported in the literature such as the Hestrin/Schramm medium describedin U.S. Pat. No. 4,378,431 (i.e., 2.0%, w/v glucose, 0.5%, w/v peptone,0.5% w/v yeast extract, 0.27%, w/v disodium phosphate, 0.11%, w/v citricacid with pH adjusted to 6.0 by dilute HCl or NaOH).

The matrix comprising bacterial cellulose is placed in a solution of ametal salt for a sufficient time period such that the metal salt isreduced to metallic form and the metal precipitates in or on the matrix.In one example embodiment, the metal salt is selected from coordinationcompounds including a transition metal complex ion, and mixturesthereof. In another example embodiment, the metal salt is selected fromcoordination compounds including a platinum metal group (i.e.,ruthenium, osmium, rhodium, iridium, platinum and palladium) complexion, and mixtures thereof. In yet another example embodiment, the metalsalt is a coordination compound including a palladium complex ion. Themetal loading in or on the bacterial cellulose matrix can be varied bycontrolling the amount of metal salt and the incubation time. Also, theco-precipitation of different metals can be achieved. After the selectedincubation time has been reached, the matrix is removed from thesolution and may be dried using a standard gel drier. The drying stepcauses the cellulose matrix to become dehydrated to a thin membrane, asthe bacterial cellulose in its native hydrated form contains 100 to 200times its weight in water.

Certain metal salts will be spontaneously precipitated by the bacterialcellulose whereas other metal salts will not be spontaneouslyprecipitated by the bacterial cellulose. It has been discovered that ifthe metal salt is a coordination compound including a transition metalcomplex ion, the metal may or may not be spontaneously precipitated bythe bacterial cellulose depending on the chemical stability of the metalsalt. For example, when the metal salt is a coordination compoundincluding a palladium complex ion, such as ammonium hexachloropalladate,spontaneous deposition of metallic palladium in the bacterial cellulosematrix diffused with ammonium hexachloropalladate is observed. Incontrast, when the metal salt is a coordination compound including aplatinum complex ion, such as ammonium hexachloroplatinate, spontaneousdeposition of metallic platinum in the bacterial cellulose matrix maynot be observed. However, the ammonium hexachloroplatinate may reducedby perfusion of hydrogen gas or other reductant into the bacterialcellulose matrix thereby allowing for precipitation of metallicplatinum. In this manner, the deposition other metals in the bacterialcellulose may also be achieved, such as the precipitation of metallicgold from hexachloroaurate compounds diffused in bacterial cellulose.

It has been discovered that the metal impregnated bacterial cellulosecan be used in a variety of applications. For example, in a secondaspect of the invention, the metal impregnated bacterial cellulose isused in the construction of a fuel cell. In this second aspect of theinvention, a hydrated bacterial cellulose support structure is placed ina solution of a metal salt to precipitate the metal as described above.Contact wires are then inserted in two pieces of the metal impregnatedsupport structure, and the two pieces of metal impregnated supportstructure are then placed on opposite sides of a layer of hydratedbacterial cellulose. The three layer structure is then dehydrated tocreate a fuel cell. The resulting fuel cell has a dehydrated metalimpregnated bacterial cellulose anode, a dehydrated bacterial celluloseelectrolyte layer, and a dehydrated metal impregnated bacterialcellulose cathode.

Various methods are available for constructing a fuel cell in accordancewith the second aspect of the invention. In a first version forconstructing a fuel cell according to the invention, an electrodesupport structure comprising hydrated bacterial cellulose is firstprepared using known methods for preparing bacterial cellulose such asthose described above. The electrode support structure is then placed ina solution of a metal salt for a sufficient time period such that themetal salt is reduced to metallic form and the metal precipitates in oron the electrode support structure. Non-limiting examples of suitablemetal salts include the coordination compounds listed above, and inparticular, coordination compounds including a platinum metal group(e.g., platinum and palladium) complex ion, and mixtures thereof arequite useful. The metal loading in or on the electrode support structurecan be varied by controlling the amount of metal salt and the incubationtime. Also, the co-precipitation of different metals can be achieved.The electrode support structure is then removed from the solution of ametal salt, and at least two contacts are placed in the electrodesupport structure. One example of a suitable contact is an electricallyconductive carrier such as a platinum wire.

The electrode support structure is then dehydrated to form an electrodematerial. One suitable instrument for dehydrating the electrode supportstructure is a standard gel drier designed to dry polyacrylamide oragarose gels. The dry electrode material is then divided into an anodeand a cathode, each of which has at least one contact. A membranesupport structure comprising hydrated bacterial cellulose is alsoprepared using known methods for preparing bacterial cellulose such asthose described above. The anode is then placed on one side of themembrane support structure, and the cathode is then placed on anopposite side of the membrane support structure, thereby creating athree layer anode—membrane support structure—cathode structure. Themembrane support structure is then dehydrated (such as by the gel drier)thereby affixing the anode and the cathode to the membrane supportstructure and forming a fuel cell. No adhesives are required to hold themulti-layered structure together as the hydrogen bonds between thecellulose fibrils are sufficient to keep the multi-layered structureintact. The resulting fuel cell has a dehydrated metal impregnatedbacterial cellulose anode, a dehydrated bacterial cellulose electrolytelayer, and a dehydrated metal impregnated bacterial cellulose cathode.When the contact wires of the fuel cell are connected to an electricalcircuit and the anode is contacted with a hydrogen-containing fuel, thefuel cell generates electricity as measured in the circuit. Inparticular, the halfcell reactions at the anode and the cathode are,respectively: H₂→2H⁺+2e⁻ and ½O₂+2H⁺+2e⁻→H₂O.

While the process steps in the first version for constructing a fuelcell according to the invention have been described in a specific order,it can be appreciated that a fuel cell can be prepared by undertakingthe process steps in other sequences and with other variations of theindividual process steps. For example, in a second version forconstructing a fuel cell according to the invention, separate first andsecond electrode support structures are prepared from bacterialcellulose and then are separately placed in a solution of a metal saltfor a sufficient time period such that the metal salt is reduced tometallic form and the metal precipitates in or on the first and secondelectrode support structures. Contact wires are then placed in eachelectrode support structure and the electrode support structures aredehydrated to form an anode and a cathode. The anode and the cathode maythen be affixed to a membrane support structure as described above inthe first version for constructing a fuel cell.

In a third version for constructing a fuel cell according to theinvention, the electrode support structure may prepared from bacterialcellulose and then placed in a solution of a metal salt for a sufficienttime period such that the metal salt is reduced to metallic form and themetal precipitates in or on the electrode support structure. Theelectrode support structure is then divided into a first portion and asecond portion. Contact wires are then placed in each portion of theelectrode support structure and the portions of electrode supportstructure are dehydrated to form an anode and a cathode. The anode andthe cathode may then be affixed to a membrane support structure asdescribed above in the first version for constructing a fuel cell.

In a fourth version for constructing a fuel cell according to theinvention, the electrode support structure may prepared from bacterialcellulose and then placed in a solution of a metal salt for a sufficienttime period such that the metal salt is reduced to metallic form and themetal precipitates in or on the electrode support structure. Theelectrode support structure is then divided into an anode portion and acathode portion. Contact wires are then placed in each portion of theelectrode support structure. The anode portion of the electrode supportstructure and a cathode portion of the electrode support structure arethen placed on opposite sides of membrane support structure as preparedabove and the anode portion, cathode portion and membrane supportstructure are simultaneously dehydrated to form a fuel cell.

In a fifth version for constructing a fuel cell according to theinvention, separate first and second electrode support structures areprepared from bacterial cellulose and then are separately placed in asolution of a metal salt for a sufficient time period such that themetal salt is reduced to metallic form and the metal precipitates in oron the first and second electrode support structures. Contact wires arethen placed in each electrode support structure. The first electrodesupport structure and the second electrode support structure are thenplaced on opposite sides of membrane support structure as prepared aboveand the first electrode support structure, the second electrode supportstructure and the membrane support structure are simultaneouslydehydrated to form a fuel cell.

It has been discovered that the electrical current generated by a fuelcell according to the invention can be increased by pretreating thelayer of bacterial cellulose forming the membrane support structure witha metal salt and/or a sulfonated polymer before dehydration. In a firstversion of this membrane pretreatment process step, a membrane supportstructure comprising hydrated bacterial cellulose is prepared usingknown methods for preparing bacterial cellulose such as those describedabove. The membrane support structure may then be placed in a solutionof a metal salt for a sufficient time period such that the metal salt isdeposited in or on the membrane support structure. Non-limiting examplesof suitable metal salts include alkali metal salts, and in particular,alkali metal chlorides such as potassium chloride are quite beneficial.In a second version of this membrane pretreatment process step, amembrane support structure comprising hydrated bacterial cellulose isplaced in a solution of a sulfonated polymer for a sufficient timeperiod such that the sulfonated polymer is deposited in or on themembrane support structure. Non-limiting examples of suitable sulfonatedpolymers include perfluorinated sulfonic acid polymers such as thosesold under trademark “Nafion”, sulfonated poly(aryl ether ketones),sulfonated polyaromatic polymers such as those described in U.S. Pat.Nos. 3,528,858 and 3,226,361, and natural sulfonated polymers such ascarrageenan. In a third version of this membrane pretreatment processstep, a membrane support structure comprising hydrated bacterialcellulose is first placed in a solution of a sulfonated polymer for asufficient time period such that the sulfonated polymer is deposited inor on the membrane support structure and then placed in a solution of ametal salt for a sufficient time period such that the metal salt isdeposited in or on the sulfonated polymer and/or the membrane supportstructure.

The above-described methods for constructing a fuel cell according tothe invention produce fuel cell electrodes and electrolyte membranesthat are also advantageous when incorporated into other fuel cellconfigurations. In other words, the bacterial cellulose based electrodesand bacterial cellulose based electrolyte membranes do not necessarilyhave to be used together in the same fuel cell but can also be used withnon-cellulose based electrodes and non-cellulose based electrolytemembranes. A fuel cell electrode is prepared according to the inventionby impregnating bacterial cellulose with a transition metal catalyst(e.g., platinum or palladium), inserting an electronically conductivecurrent carrier (such as a platinum group metal wire) into the bacterialcellulose, and dehydrating the bacterial cellulose. Co-precipitation ofdifferent transition metal catalysts can also be used to vary thecatalytic properties of the electrode. A fuel cell electrode prepared inthis manner has a current carrier (e.g., wire contact), a catalyst(e.g., palladium), and a proton transferring substrate (bacterialcellulose) and therefore, would be suitable for use as an anode and/orcathode with conventional solid electrolyte membranes. A fuel cellelectrode prepared according to the invention is particularlyadvantageous as the catalyst can be recovered from the fuel cellelectrode by burning or hydrolyzing away the cellulose usingconventional methods and equipment. A fuel cell electrode preparedaccording to the invention using palladium also has advantageoushydrogen storage capability, and therefore has applications along theselines.

An electrolyte membrane is prepared according to the invention byimpregnating bacterial cellulose with a metal salt (e.g., potassiumchloride) and/or a sulfonated polymer (e.g., carrageenan) anddehydrating the bacterial cellulose. Typically, when a metal salt and asulfonated polymer are used, the sulfonated polymer is infusedthroughout the cellulose, followed by soaking in the metal salt to loadelectrolyte ions on the sulfonate groups of the sulfonated polymer. Anelectrolyte membrane prepared in this manner has a proton transferringsubstrate (bacterial cellulose) and additional electrolytes (a metalsalt and/or a sulfonated polymer) and therefore, would be suitable foruse with conventional electrodes.

A fuel cell electrode prepared according to the invention can be furtherprocessed to produce an enzyme electrode. An enzyme electrode comprises,in most cases, an enzyme immobilized on an electrode as a base. When anoxidizable substance (e.g., glucose, lactose, cholesterol) is contactedwith the enzyme (e.g., glucose oxidase, lactate oxidase, or cholesteroloxidase), an enzymatic reaction proceeds which gives an electrodecurrent, and the electrode current changes depending on the amount ofthe oxidizable substance. The concentration of the oxidizable substanceis determined by the change of the electrode current with reference to apreviously formed calibration curve. Therefore, an enzyme electrodeaccording to the invention can be prepared by depositing an enzyme in oron a fuel cell as prepared above.

EXAMPLES

The following examples are intended only to further illustrate theinvention and are not intended to limit the scope of the invention whichis defined by the claims.

Example 1 Preparation of Bacterial Cellulose

The bacterial cellulose used in the process was the Philippine foodproduct nata de coco that is produced from coconut milk and sucrose byfermentation with a starter culture containing the bacteriumGluconoacetobacter xylinus (formerly Acetobacter xylinum) or relatedspecies. (In an alternative method, a similar cellulose product can beproduced by growing G. xylinus on media formulations that have beenreported in the literature such as the Hestrin/Schramm medium describedin U.S. Pat. No. 4,378,431).

Example 2 Incubation of Bacterial Cellulose Cubes with Palladium

Bacterial cellulose cubes were incubated with ammoniumhexachloropalladate dissolved in 50 mM sodium acetate pH 4.5 at 38° C.This resulted in the precipitation of palladium metal throughout thebacterial cellulose cubes. The palladium metal precipitation beganwithin 30 minutes of the start of incubation and was allowed to continuefor 18 to 24 hours, after which time the cubes were completely black inappearance. This material, consisting of cellulose impregnated withpalladium particles, is referred to herein as “palladium-cellulose”.Analysis of the hexachloropalladate ion remaining in solution indicatedthat 30% of total added had precipitated in or on the bacterialcellulose. Incubation of the bacterial cellulose cubes in a solution ofammonium hexachloropalladate dissolved in HPLC-grade water increased theamount of palladium precipitated in the cubes to 50% of the totalhexachloropalladate added. The amount of palladium in the cellulosecubes increased as the concentration was increased from 5 to 10 mM, butthe fraction of palladium precipitated per hexachloropalladate added wasthe same for each solution. Reduction of the hexachloropalladate metalcomplex to metal particles occurred spontaneously. Without intending tobe bound by theory, it appears that the reduction of thehexachloropalladate metal complex to metal particles is carried out bythe reducing ends of the cellulose chains comprising the bacterialcellulose.

Example 3 Incubation of Bacterial Cellulose Cubes with Platinum

For platinization of the cellulose, ammonium hexachloroplatinate isfirst loaded into the cubes by diffusion during overnight incubation asdescribed for palladium in Example 2. The hexachloroplatinate is notspontaneously reduced inside the cellulose, but is reduced by perfusionof hydrogen gas (or other reductant) into the cellulose matrix. Becausehexachloroplatinate is more stable than the corresponding palladiumsalt, it is believed that this makes it more resistant to reduction bythe reducing ends of the cellulose fibrils. Precipitation ofhexachloroaurate has also been demonstrated in bacterial cellulose.

Example 4 Dehydration of Metal Impregnated Bacterial Cellulose Cubes

The cubes of cellulose containing precipitated metal were rapidly driedunder vacuum using a standard gel drier designed to dry polyacrylamideor agarose gels. The drying step caused the cellulose cubes or sheets tobecome dehydrated to a thin membrane, as the bacterial cellulose in itsnative hydrated form contains 100 to 200 times its weight in water. Forassembly of multilayer devices, the pretreated cubes are driedsequentially, by placing a hydrated cube on top of the previously driedcube, then applying vacuum. No adhesives are required to hold themulti-layered structures together. The hydrogen bonds between thecellulose fibrils are sufficient to keep the multi-layered structureintact.

Example 5 Construction of a Membrane Electrode Assembly

A membrane electrode assembly suitable for use in a fuel cell wasconstructed by layering catalyst and insulator layers. Thepalladium-cellulose layers act as the catalyst for the twohalf-reactions of the fuel cell. To prepare an insulating layer, a cubeof untreated bacterial cellulose was dehydrated on the gel dryer for 30seconds to dry to a thin membrane. Catalyst membranes are prepared byinsertion of platinum wires into a hydrated metallized cube beforedrying. The construction of the catalyst membranes is schematicallyshown in FIGS. 1A and 1B. To prepare a catalyst layer, four platinumwires (two anode wires 13 in FIGS. 1A and 1B and two cathode wires 16 inFIGS. 1A and 1B) with a diameter of 0.1 millimeters were inserted into acube of palladium-cellulose at regular intervals. Thepalladium-cellulose cube catalyst layer with the inserted wires wasplaced on top of the insulating layer and the drying process wasrepeated. The untreated insulating layer of bacterial cellulose servesto protect the palladium-cellulose layer from the external environment.The resulting structure was trimmed along the edges to ensure that thecatalyst and insulating layers were identical in size. This layeredmembrane assembly was then cut in half, so that each half contained twoplatinum wires. The layered membranes are depicted in FIGS. 1A and 1B.The dashed line in FIGS. 1A and 1B indicates a parting line between ananode 12 and an anode protective layer 11, and a cathode 15 and acathode protective layer 17. These two pieces are the cathode and anodeof the fuel cell. A layered membrane electrode assembly is shown in FIG.2. Looking at FIG. 2, two layers of unmodified bacterial cellulose(element 14 in FIG. 2) were used to separate the catalyst layers in afuel cell 10, as cellulose is a good insulator and prevents electronflow between the catalyst layers. Protons can permeate through thecellulose layers, and the platinum wires complete the circuit. Anadditional layer of bacterial cellulose was added to protect thecatalyst layer from the environment.

Example 6 Assembly of Fuel Cell from the Membrane Electrode Assembly

The membrane electrode assembly was used as part of a fuel cell that wasplaced in a small hydrogen generator assembly to test for currentproduction. The assembly is depicted in FIG. 3. The assembly includestwo glass fittings 29 a and 29 b with O-ring joints 27 a and 27 b. Thefuel cell 10 of FIG. 2 was sandwiched between two O-rings 26 that alsocontact the O-ring joints 27 a and 27 b of the glass fittings 29 a and29 b. A reaction vessel 28 including O-ring joint 27 c was fittedagainst O-ring joint 27 d of the bottom glass fitting 29 b. Hydrogen wasgenerated in the reaction vessel 28 using metal displacement orelectrolysis. The platinum wires 13 and 16 were attached to anautoranging picoammeter that was interfaced with a computer running adata collection program sold under the trademark “Labview” forcontinuous on-line monitoring of current production. Current productionwas monitored for at least 24 hours to test each fuel cellconfiguration. In FIG. 4, there is shown a plot of current production(electrical production) in microamps versus time (hours) for the fuelcell configuration having untreated dehydrated bacterial cellulose asthe electrolyte layer and acid displacement (Fe/acetic acid reaction) asthe hydrogen source. A control experiment was also carried out toconfirm that the current measured was not a result of H₂ oxidation onthe platinum wires.

Example 7 Construction of an Alternative Membrane Electrode Assembly

An alternative electrolyte layer was prepared in an effort to increasethe performance of the fuel cell. As in Example 5, a membrane electrodeassembly suitable for use in a fuel cell was constructed by layeringcatalyst and insulator layers. The palladium-cellulose layers act as thecatalyst for the two half-reactions of the fuel cell. The insulatinglayer was prepared by dehydrating a cube of untreated bacterialcellulose on a gel dryer for 30 seconds to dry to a thin membrane.Catalyst membranes were prepared by insertion of platinum wires into ahydrated metallized cube before drying. The catalyst layer was preparedby inserting four platinum wires with a diameter of 0.1 millimeters intoa cube of palladium-cellulose at regular intervals. Thepalladium-cellulose cube catalyst layer with the inserted wires wasplaced on top of the insulating layer and the drying process wasrepeated. The untreated insulating layer of bacterial cellulose servesto protect the palladium-cellulose layer from the external environment.The resulting structure was trimmed along the edges to ensure that thecatalyst and insulating layers were identical in size. This layeredmembrane assembly was then cut in half, so that each half contained twoplatinum wires. The layered membranes are depicted in FIGS. 1A and 1B.These two pieces are the cathode layer and the anode layer of the fuelcell.

An electrolyte layer was then prepared by infusing a cube of untreatedbacterial cellulose with 1 M potassium chloride (KCl) for at least 24hours. The KCl-treated bacterial cellulose was then dried between theanode and cathode layers for 10 minutes on the gel-dryer to create amembrane electrode assembly. The physical characteristics of the fuelcell were as follows in Table 1.

TABLE 1 Surface area 4.0 cm² Area of Catalyst layer 1.53 cm²  Area ofCatalyst layer exposed to H₂ 1.1 cm² Palladium loading 1 mg/catalystlayer Surface area of electrodes in 0.05 cm²  contact with catalystWeight of fuel cell 126 mg  (Approx. weight of Pt wires is 100 mg)Operating temperature 26° C.

The membrane electrode assembly was used as part of a fuel cell that wasplaced in a small hydrogen generator assembly to test for currentproduction as in Example 6 above. Current production was monitored forat least 24 hours to test the fuel cell configuration. The performanceof the fuel cell was tested using three different H₂ sources: aciddisplacement reaction, a 4% H₂/96% Ar gas mixture, and 100% H₂. Opencircuit current and voltage measurements of the fuel cells were takenand are shown in Table 2.

TABLE 2 Current Voltage H₂ source Reaction Conditions (μA) (V) Aciddisplacement Fe/acetic acid reaction. The 192 0.483 reaction was underpressure due to the build up of H₂ in the vessel 4% H₂/96% Aratmospheric pressure, 25 0.442 71.6 mmol H₂/min 100% H₂ atmosphericpressure, 36 0.273 1.78 mol H₂/min

In FIG. 5, there is shown a plot of current production (electricalproduction) in microamps versus time (hours) for the fuel cellconfiguration having KCl-treated dehydrated bacterial cellulose as theelectrolyte layer and acid displacement (Fe/acetic acid reaction) as thehydrogen source. Voltage and current values for the fuel cell using the4% H₂/96% Ar gas mixture and the fuel cell using 100% H₂ were alsomeasured. In this test, the resistance was increased in 2 kilo-ohmincrements from 1 kilo-ohm to 11 kilo-ohm. The testing was carried outat 28° C. and atmospheric pressure, and the gas flow rate was 40milliliters per minute. The plot of voltage versus current for the fuelcell using the 4% H₂/96% Ar gas mixture and the fuel cell using 100% H₂are shown in the graph of FIG. 6.

Example 8 Construction of Another Alternative Membrane ElectrodeAssembly

Another alternative electrolyte layer was prepared in an effort toincrease the performance of the fuel cell. This type of electrolytemembrane was prepared by first soaking the bacterial cellulose inK-carrageenan to infuse the sulfated polysaccharide throughout thecellulose layer, followed by soaking in potassium chloride to loadelectrolyte ions on the sulfonate groups of the K-carrageenan. Thismembrane was used as the electrolyte membrane in a fuel cellconstruction in the same manner as the KCl-impregnated cellulosedescribed above in Example 7. The fuel cells constructed with thepotassium chloride-carrageenan membranes were tested as in Example 7 andgave test results in the production of current from hydrogen similar tothose obtained with the KCl-cellulose membrane.

Thus, there has been provided a method for the deposition of metals inbacterial cellulose and for the employment of the metallized bacterialcellulose in the construction of fuel cells and other electronicdevices. The methods have several advantages. First, metal salts can beinfused into the bacterial cellulose matrix, reduced to metallic form,precipitated inside the cellulose, and then concentrated by dryingwithout the use of toxic solvents or caustic solutions. Second, themetal loading on the bacterial cellulose matrix can be varied bycontrolling the amount of metal salt and incubation time. Third, thebacterial cellulose is capable of absorbing small inorganic moleculesand large bio-molecules, and therefore, it is easy to alter theproperties of the bacterial cellulose by infusing in the desired reagentprior to drying. For example, proton or electronic conducting propertiescan be conferred on the bacterial cellulose by addition of theappropriate electrolyte, and the properties of the bacterial cellulosecan also be changed by chemical modification of functional groups on thecellulose fibrils, such as by sulfonation of the bacterial cellulose toproduce a material with proton conducting properties. Fourth, assemblyof a fuel cell can be achieved by sequential drying of treated and/oruntreated cellulose layers, with no requirement for glues. Fifth, theuntreated and treated bacterial cellulose is stable to autoclaving (120°C. and 20 psi.), and therefore the untreated and treated bacterialcellulose is a versatile material that can be used for low temperature(20-80° C.) and mid range temperature (80-150° C.) fuel cells. Sixth,for fuel cell applications, the low cost, lightweight, and low toxicityof the dehydrated untreated and treated bacterial cellulose are seen asmajor advantages compared to other methods. Seventh, the recovery of thecatalyst (e.g., palladium) from the fuel cell electrodes and membranesis quite simple, as the cellulose portion can be burned or hydrolyzedaway from the metals using conventional methods and equipment. Otheradvantages would be apparent to those skilled in the art.

Although the present invention has been described in considerable detailwith reference to certain embodiments, one skilled in the art willappreciate that the present invention can be practiced by other than thedescribed embodiments, which have been presented for purposes ofillustration and not of limitation. Therefore, the scope of the appendedclaims should not be limited to the description of the embodimentscontained herein.

1. A method for forming a fuel cell, the method comprising: providing anelectrode support structure comprising bacterial cellulose; placing theelectrode support structure in a solution of a metal salt for asufficient time period such that the metal salt is reduced to metallicform and the metal precipitates in or on the electrode support structureto form an electrode material; providing an electrolyte membrane;placing an anode on one side of the electrolyte membrane; and placing acathode on an opposite side of the electrolyte membrane, wherein one ofthe anode and the cathode comprises the electrode material.
 2. Themethod of claim 1 wherein: the anode comprises the electrode materialand the cathode comprises the electrode material.
 3. The method of claim1 wherein: the electrode support structure comprises hydrated bacterialcellulose, and the method further comprises dehydrating the electrodesupport structure after the metal precipitates in or on the electrodesupport structure to form the electrode material.
 4. The method of claim1 wherein: the electrolyte membrane comprises bacterial cellulose. 5.The method of claim 1 wherein: the electrolyte membrane compriseshydrated bacterial cellulose, the electrode support structure compriseshydrated bacterial cellulose, the anode comprises the electrodematerial, the cathode comprises the electrode material, and the methodfurther comprises dehydrating the electrolyte membrane, the anode andthe cathode after placing the anode on one side of the electrolytemembrane and placing the cathode on the opposite side of the electrolytemembrane thereby affixing the anode and the cathode to the electrolytemembrane.
 6. The method of claim 1 wherein: the electrolyte membranecomprises hydrated bacterial cellulose, the electrode support structurecomprises hydrated bacterial cellulose, the method further comprisesdehydrating the electrode support structure after the metal precipitatesin or on the electrode support structure to form the electrode material,the anode comprises the electrode material, the cathode comprises theelectrode material, and the method further comprises dehydrating theelectrolyte membrane after placing the anode on one side of theelectrolyte membrane and placing the cathode on the opposite side of theelectrolyte membrane thereby affixing the anode and the cathode to theelectrolyte membrane.
 7. The method of claim 1 wherein: the metal saltis selected from coordination compounds including a platinum metal groupcomplex ion, and mixtures thereof.
 8. The method of claim 1 wherein themetal salt is a coordination compound including a palladium complex ion.9. The method of claim 1 wherein the method further comprises contactingthe electrode support structure with a reductant fluid when theelectrode support structure is placed in the solution of the metal salt.10. The method of claim 1 wherein the electrolyte membrane comprisesbacterial cellulose and a sulfonated polymer.
 11. A method forrecovering the metal from the fuel cell produced by the method of claim1, the method for recovering comprising burning or hydrolyzing theelectrode material.
 12. A method for forming an electrolyte membrane fora fuel cell, the method comprising: providing a support structurecomprising hydrated bacterial cellulose; infusing the support structurewith a metal salt and/or a sulfonated polymer; and dehydrating thesupport structure to form the electrolyte membrane.
 13. The method ofclaim 12 wherein: the metal salt is selected from alkali metal salts.14. The method of claim 12 wherein: the support structure is infusedwith a metal salt and a sulfonated polymer.
 15. The method of claim 12wherein: the support structure is infused with a metal salt.
 16. Themethod of claim 12 wherein: the support structure is infused with asulfonated polymer.
 17. An electrolyte membrane for a fuel cell, theelectrolyte membrane comprising: a support structure comprisingbacterial cellulose; and a metal salt disposed in or on the supportstructure.
 18. The electrolyte membrane of claim 17 wherein: the metalsalt is selected from alkali metal salts.
 19. The electrolyte membraneof claim 17 further comprising: a sulfonated polymer disposed in or onthe support structure.
 20. The electrolyte membrane of claim 19 wherein:the sulfonated polymer is carrageenan.